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Objective: Existing methods to evaluate skin care products suffer limitations. This is the case for ex vivo skin explants, a first-choice 3D model. While essential to analyse mid- to long-term biological effects, this classical model hinders assessing microrelief variations. To circumvent these limitations, we developed an ex vivo PERFused EXplant setup (Perfex) that maintains the outer skin surface in the open air, closely mirroring physiological conditions., Methods: A custom-designed reservoir enables perfusing the dermal side of explants with buffered, temperature-controlled medium, while the epidermis is subjected to "normal" conditions. Skin tension and characteristics of the stratum corneum, microrelief, histology and immunohistology (collagen types I and III, elastin and fibrillin-1) were analysed and compared to those of explants maintained under conventional conditions or in vivo skin. The effects of skin care formulas intended to induce short- and/or mid- to long-term effects were also assessed., Results: Skin explants maintained with the Perfex setup exhibit characteristics (firmness, elasticity, hydration and barrier function) closer to those of in vivo skin than with conventional conditions. Moreover, Perfex-maintained explants present no alteration in histology after 7 days and slight variation in the expression of key protein markers. Microrelief characteristics also remain mostly stable over 7 days. Formula applications corroborate that skin tensor-containing products primarily induce short-term changes in the microrelief, while those with biologically active ingredients mainly lead to mid- to long-term effects on the histology and expression of molecular markers. Furthermore, maintaining skin explants with a physiologically relevant skin surface enabled analysing the relationship between microrelief and key markers, showing that fibrillin-1 is the protein most correlated with microrelief characteristics., Conclusions: The Perfex setup allows for similar preservation of skin explant histology and key protein expression as the conventional system, yet it maintains a skin surface close to that of in vivo skin. Therefore, it is valuable to analyse both the short- and mid- to long-term impacts of skin care formulas and better comprehend their effects. The Perfex system also offers a new tool for investigating fundamental questions, such as the link that can exist between dermal proteins and skin surface properties., (© 2024 Society of Cosmetic Scientists and Societe Francaise de Cosmetologie.)

Air pollution is a growing threat to human health. Airborne pollution effects on respiratory, cardiovascular and skin health are well-established. The main mechanisms of air-pollution-induced health effects involve oxidative stress and inflammation. The present study evaluates the potential of a polyphenol-enriched food supplement ingredient comprising Lippia citriodora , Olea europaea , Rosmarinus officinalis , and Sophora japonica extracts in mitigating the adverse effects of environmental pollution on skin and cardiopulmonary systems. Both in vitro and ex vivo studies were used to assess the blend's effects against pollution-induced damage. In these studies, the botanical blend was found to reduce lipid peroxidation, inflammation (by reducing IL-1α), and metabolic alterations (by regulating MT-1H, AhR, and Nrf2 expression) in human skin explants exposed to a mixture of pollutants. Similar results were also observed in keratinocytes exposed to urban dust. Moreover, the ingredient significantly reduced pollutant-induced ROS production in human endothelial cells and lung fibroblasts, while downregulating the expression of apoptotic genes (bcl-2 and bax) in lung fibroblasts. Additionally, the blend counteracted the effect of urban dust on the heart rate in zebrafish embryos. These results support the potential use of this supplement as an adjuvant method to reduce the impact of environmental pollution on the skin, lungs, and cardiovascular tissues.

Photobiomodulation (PBM) is rapidly gaining traction as a valuable tool in dermatology for treating many inflammatory skin conditions using low levels of visible light or near-infrared radiation. However, the physiological regulatory pathways responsible for the anti-inflammatory effect of PBM have not been well defined. Since previous studies showed that nuclear factor-erythroid 2 like 2 (Nrf2) is a master regulator of the skin inflammatory response, we have addressed its role in controlling inflammation by PBM. Primary human keratinocytes (KCs) stimulated with 2,4-dinitrochlorobenzene (DNCB) to mimic pro-inflammatory stress were illuminated with two wavelengths: 660 nm or 520 nm. Both lights significantly reduced the mRNA expression of the DNCB-triggered TNF-α , IL-6 , and IL-8 cytokines in KCs, while they enhanced Nrf2 pathway activation. PBM-induced Nrf2 is a key regulator of the inflammatory response in KCs since its absence abolished the regulatory effect of light on cytokines production. Further investigations of the mechanisms contributing to the immunoregulatory effect of PBM in inflamed human skin explants showed that 660 nm light prevented Langerhans cells migration into the dermis, preserving their dendricity, and decreased pro-inflammatory cytokine production compared to the DNCB-treated group. This study is the first to report that the PBM-mediated anti-inflammatory response in KCs is Nrf2-dependent and further support the role of PBM in skin immunomodulation. Therefore, PBM should be considered a promising alternative or complementary therapeutic approach for treating skin-related inflammatory diseases.

The skin epidermis is continuously exposed to external aggressions, including environmental pollution. The cosmetic industry must be able to offer dedicated products to fight the effects of pollutants on the skin. We set up an experimental model that exposed skin explants maintained in culture to a pollutant mixture. This mixture P representing urban pollution was designed on the basis of the French organization 'Air Parif' database. A chamber, called Pollubox®, was built to allow a controlled nebulization of P on the cultured human skin explants. We investigated ultrastructural morphology by transmission electron microscopy of high pressure frozen skin explants. A global transcriptomic analysis indicated that the pollutant mixture was able to induce relevant xenobiotic and antioxidant responses. Modulated detoxifying genes were further investigated by laser micro-dissection coupled to qPCR, and immunochemistry. Both approaches showed that P exposure correlated with overexpression of detoxifying genes and provoked skin physiological alterations down to the stratum basale. The model developed herein might be an efficient tool to study the effects of pollutants on skin as well as a powerful testing method to evaluate the efficacy of cosmetic products against pollution., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objective: The aim of the paper presented herein is the description and safety evaluation of the process of dissolution of an 86-microneedle patch composed of hyaluronic acid, when applied topically to human abdominal skin explants. Such explants were chosen to replace the inability of obtaining periorbital skin. In order to evaluate penetration and dissolution of the microneedles, we employed histochemical methods and a fluorescent dye FITC (fluorescein isothiocyanate)., Methods: Abdominoplasty human skin explants were treated with square microneedle patches with a 1.5-cm 2 surface area, containing 86 microneedles and having 450 ± 23.5 µm in height with 1 mm interspacing between nearest neighbouring microneedles. Histological processing and staining for cell viability, FITC distributions and glycosaminoglycans were performed. The stained surface percentage for each treatment was compared to control untreated samples at given time points. A Mann-Whitney test was used to identify the difference between two populations (sites of skin samples punctured with stained and clear microneedles, respectively) at the given level of statistical significance (P < 0.05)., Results: The application of the MN patch to excised skin explants showed these microneedles to be non-invasive into the dermis of the skin. Skin puncturing with MN patches revealed 17 different sites of microneedle penetration immediately afterwards and 4 sites, 2 h later. Although there were some variances in the epidermal depth of penetration, these variances did not impact on cell viability. The hyaluronic acid-based microneedles having 450 µm in length penetrated the epidermis at an averaged depth by 26 µm without disrupting skin cell viability and without causing an inflammatory response. Hyaluronic acid could be detected in most of these penetration sites, with no diffusion into the dermis, which is important for cosmetic applications. FITC analysis uncovered fluorescein isothiocyanate distribution within microneedle insertion site, which remained steady after 2 and 6 h of experimentation., Conclusion: Using ex vivo tracer staining studies, we have shown that the evaluated microneedle applicator is capable of penetrating the skin epidermis and delivering substances embedded in the needle polymer matrix. In addition, the tested product was shown to be safe, which provides a broad perspective for delivering cosmetic and pharmaceutic agents., (© 2020 Society of Cosmetic Scientists and the Société Française de Cosmétologie.)

Background: Accumulation of advanced glycation end-products (AGEs) in skin has been associated with skin aging. Inhibition of glycation of proteins of extracellular matrix may help skin texture and appearance. The objective of the study was to demonstrate the antiglycation activity of topically applied carnosine and novel facial cream (FC) containing carnosine in human skin explants ex vivo., Methods: Glycation was induced in human skin explants by methylglyoxal (MG) in culture media. FC containing carnosine (FC-CARN) or carnosine in aqueous solution (AQ-CARN) was applied topically on skin explants. Levels of AGEs carboxymethyl-lysine (CML) and pentosidine were determined in the epidermis and dermis of skin sections and were used to calculate antiglycation activity., Results: Exposure to MG led to increases in CML and pentosidine in skin explants. Antiglycation effect for AQ-CARN was CML: -64 and -41%, pentosidine: -48 and 42% in epidermis and reticular dermis respectively. Antiglycation effect for FC-CARN was CML: -150 and -122%, pentosidine: -108 and -136%, in epidermis and reticular dermis respectively., Conclusion: Topically applied carnosine protects against the glycation induced by MG. Novel FC-CARN significantly reduced levels of AGEs in both epidermis and reticular dermis in human skin explants., (© 2018 S. Karger AG, Basel.)

Background: Hyaluronic acid (HA) is a popular ingredient in topical formulations for cosmetic improvement of the skin. Most formulations contain linear, non-crosslinked HA oligomers, low molecular weight (LMW) HA, and/or high molecular weight (HMW) HA. Crosslinking of HA enhances its clinical longevity and mechanical characteristics. The objective of this study was to characterize the topical effects of a new, crosslinked resilient HA (RHA) that is also available as a cohesive, tissue-integrating injectable filler, compared with non-crosslinked HMW HA and LMW HA. Living human skin explants that preserve the 3-dimensional structure of in vivo skin were used to maximize clinical relevance., Methods: Standardized doses of each HA product were applied daily for 9 days to human skin explant surfaces. Untreated explants served as controls. Water content of the stratum corneum and entire epidermis was analyzed by Raman spectroscopy. Transepidermal water loss (TEWL) was measured to assess skin barrier function. Explant morphology and microrelief were evaluated by optical and scanning electron microscopy., Results: Crosslinked RHA achieved a significant increase in epidermal water content (7.6%) over the control. Spectral cartography confirmed a higher epidermal water content with RHA than with HMW HA or LMW HA. TEWL was reduced by 27.8% with RHA, and by 15.6% with HMW HA, but increased by 55.5% with LMW HA. Cutaneous microrelief improved with RHA. Corneocyte cohesion improved with RHA and HMW HA., Conclusions: This comparative, multimodal study demonstrated greater benefits of topical crosslinked RHA over linear HMW HA or LMW HA in reducing TEWL, retaining and redistributing water within the epidermis, maintaining skin integrity, and improving skin barrier structure and function. RHA was a more efficacious humectant than LMW HA, and a more efficacious occlusive moisturizer than HMW HA. These integrative epidermal repair activities are of significant value for addressing primary deficits of aging skin, improving tolerance to retinoids and other topical agents, and optimizing procedural outcomes. A combination of topical and injectable HA provides an elegant model of synergistic, multi-level skin restoration.

Glycation is an ageing reaction of naturally occurring sugars with dermal proteins, whose clinical signs may appear in vivo around age 30, and increases steadily/regularly with age. The suppleness of the dermis is affected by the formation of bridges between proteins and sugars (Maillard's reaction). The residues formed (Amadori products, Advanced Glycation End products) as well as the proteins they alter, can be visualized by specific immunostainings. Induced in a few days on living skin explants by methylglyoxal, glycation can be prevented by the application of aminoguanidine HCl, the reference anti-glycation molecule. This model enabled to highlight the anti-glycation activity of substances of vegetal origin such as puerarin and chlorogenic acid., (© 2011 The Authors. ICS © 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie.)

Background: Hydrofluoric acid (HF) is a small and partially dissociated acid (pK(a) 3.2), able to deeply penetrate into human skin in addition to the corrosiveness of the hydrogen ion (H(+)) and the toxicity of the fluoride ion (F(-)). However, there has been a lack of experimental studies to objectively characterize the results of human HF skin exposure decontamination., Methodology/principal Findings: A previously established experimental method using a human skin explants ex vivo model (Part 1. Experimental 70% hydrofluoric acid (HF) burns: Histological observations in an established human skin explants ex vivo model) described the lesions that appeared following 70% HF penetration. Within 5 min, 70% HF penetrates to the dermis. Using the same experimental conditions, a comparison study of two different washing protocols was performed: water + topical calcium gluconate (CaG) versus Hexafluorine(®). In these conditions, washing for 15 min with running tap water followed by topical CaG ointment only delayed burn onset, while severe tissue damage appeared later. In contrast, after washing with Hexafluorine(®) over 10 min, no histological lesions developed. These results are in accordance with the results of accidental human industrial case reports., Conclusion/significance: Amphoteric and hypertonic Hexafluorine(®) can deactivate H(+) and chelate F(-) ions. Based on these results, it should be considered as a promising first-aid decontamination solution to prevent or minimize significant local and systemic consequences of concentrated HF skin exposures.

Background: Hydrofluoric acid (HF) is particularly dangerous due to the potential for systemic effects and induction of severe skin necrosis through two mechanisms: corrosiveness and local tissue toxicity. In addition, because it is only partially dissociated (pK(a) 3.2), it is capable of penetrating deeply into tissues. There is a lack of experimental studies that objectively characterize the behavior of HF diffusion into human skin, specifically the kinetics of tissue penetration resulting in severe cellular lesions., Methodology/principal Findings: We describe the cutaneous effects of HF using an established ex vivo human skin model. The diffusion of 70% HF starts within the first minute of contact at the epidermal surface and after 2 min reaches the basal layer. In the subsequent minute, the epidermis is destroyed and lesions appear in the papillary dermis after 4 min. Soon after, damage appears in the upper reticular dermis. Thus, 70% HF needs only 5 min of contact to completely penetrate human skin explants. This experiment is reproducible and corroborates previous studies and clinical effects reported in accidental HF exposures., Conclusion/significance: This study shows that the management of HF chemical skin exposure is a question of minutes, especially for initial decontamination. These experimental observations could be useful for objectively comparing skin decontamination methods. Further studies should help to confirm these preliminary results.

Polyphenols in general are compounds that are known to promote health and have a preventive effect against various chronic diseases. The influence of cocoa polyphenols on skin, however, has scarcely been studied from a histological point of view. The aim of this study is to assess the influence of cocoa polyphenols on several indicators of skin elasticity and skin tonus, namely, glycosaminoglycans and collagen I, III and IV. This was carried out by using a model of ex vivo human skin explants maintained in survival, on which a cocoa polyphenol extract was applied. After processing by standard histological techniques (fixation, paraffin embedding, sectioning, staining, immunostaining and microscopical observation), the influence of cocoa polyphenols on the evaluated parameters was quantified by image analysis. The results obtained show that cocoa polyphenols exhibit a positive action on the parameters assessed, and the dose at which they improve the most parameters associated with skin tonus and elasticity was determined. Their activity was compared with a commercially available product, and the results obtained show that their efficacy is equivalent. Moreover, an enhancing effect of cocoa butter on activity of cocoa polyphenol was highlighted. Now that the properties of cocoa polyphenols on ex vivo skin restructuring parameters have been assessed, the next step could include their evaluation in vivo.

In a normal and healthy skin, the regular elimination of the superficial corneocytes, called desquamation, is a fundamental physiologic process intended to protect the barrier function of the skin. This invisible loss of corneocytes, individually or in small groups, is incessantly compensated by the divisions of the proliferative layer and the upward cellular maturation in order to maintain the harmonious renewal of the epidermis and the integrity of the stratum corneum. The harmony of this desquamation process is intimately conditioned by a sufficient hydration of the stratum corneum: (i) an abnormal desquamation leads to a disruption of the water barrier function and consequently to a dehydration tendency of the stratum corneum, and (ii) a cutaneous dryness (whatever the cause) is able to disturb the desquamation process. Protecting the water content of the stratum corneum has always been a major preoccupation of the cosmetic industry scientists. Consequently, the moisturizing properties of a cosmetic product are objectively measured by various explorations directly targeted on the hydration (corneometry) and on the level of the water barrier function (transepidermal water loss (TEWL) measurements), which depends directly on the skin hydration state. This intimate linkage of the desquamation process and the water content of the stratum corneum enable us to suggest an indirect assessment of the hydration from a direct study of the desquamation by examining a skin-stripping sample (D-Squames) by an optical microscope (linked to a computer). We will describe this already known technique and mainly its new and unpublished semiologic exploitation, named Diagnoskin, whose advantages are its simplicity and its reproducibility particularly interesting in the case of sequential appraisal of dermatologic or cosmetic treatments.